Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing
Fanconi anemia (FA) is a rare inherited disease characterized by developmental defects, short stature, bone marrow failure, and a high risk of malignancies. FA is heterogeneous: 15 genetic subtypes have been distinguished so far. A clinical diagnosis of FA needs to be confirmed by testing cells for...
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| Format: | Article |
| Language: | English |
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Wiley
2012-01-01
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| Series: | Anemia |
| Online Access: | http://dx.doi.org/10.1155/2012/603253 |
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| author | Johan J. P. Gille Karijn Floor Lianne Kerkhoven Najim Ameziane Hans Joenje Johan P. de Winter |
| author_facet | Johan J. P. Gille Karijn Floor Lianne Kerkhoven Najim Ameziane Hans Joenje Johan P. de Winter |
| author_sort | Johan J. P. Gille |
| collection | DOAJ |
| description | Fanconi anemia (FA) is a rare inherited disease characterized by developmental defects, short stature, bone marrow failure, and a high risk of malignancies. FA is heterogeneous: 15 genetic subtypes have been distinguished so far. A clinical diagnosis of FA needs to be confirmed by testing cells for sensitivity to cross-linking agents in a chromosomal breakage test. As a second step, DNA testing can be employed to elucidate the genetic subtype of the patient and to identify the familial mutations. This knowledge allows preimplantation genetic diagnosis (PGD) and enables prenatal DNA testing in future pregnancies. Although simultaneous testing of all FA genes by next generation sequencing will be possible in the near future, this technique will not be available immediately for all laboratories. In addition, in populations with strong founder mutations, a limited test using Sanger sequencing and MLPA will be a cost-effective alternative. We describe a strategy and optimized conditions for the screening of FANCA, FANCB, FANCC, FANCE, FANCF, and FANCG and present the results obtained in a cohort of 54 patients referred to our diagnostic service since 2008. In addition, the follow up with respect to genetic counseling and carrier screening in the families is discussed. |
| format | Article |
| id | doaj-art-b36a91fae82a4b60b494f646fd47d5d4 |
| institution | Kabale University |
| issn | 2090-1267 2090-1275 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Anemia |
| spelling | doaj-art-b36a91fae82a4b60b494f646fd47d5d42025-02-03T05:43:56ZengWileyAnemia2090-12672090-12752012-01-01201210.1155/2012/603253603253Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger SequencingJohan J. P. Gille0Karijn Floor1Lianne Kerkhoven2Najim Ameziane3Hans Joenje4Johan P. de Winter5Department of Clinical Genetics, VU University Medical Center, Van der Boechorsttraat 7, 1081 BT Amsterdam, The NetherlandsDepartment of Clinical Genetics, VU University Medical Center, Van der Boechorsttraat 7, 1081 BT Amsterdam, The NetherlandsDepartment of Clinical Genetics, VU University Medical Center, Van der Boechorsttraat 7, 1081 BT Amsterdam, The NetherlandsDepartment of Clinical Genetics, VU University Medical Center, Van der Boechorsttraat 7, 1081 BT Amsterdam, The NetherlandsDepartment of Clinical Genetics, VU University Medical Center, Van der Boechorsttraat 7, 1081 BT Amsterdam, The NetherlandsDepartment of Clinical Genetics, VU University Medical Center, Van der Boechorsttraat 7, 1081 BT Amsterdam, The NetherlandsFanconi anemia (FA) is a rare inherited disease characterized by developmental defects, short stature, bone marrow failure, and a high risk of malignancies. FA is heterogeneous: 15 genetic subtypes have been distinguished so far. A clinical diagnosis of FA needs to be confirmed by testing cells for sensitivity to cross-linking agents in a chromosomal breakage test. As a second step, DNA testing can be employed to elucidate the genetic subtype of the patient and to identify the familial mutations. This knowledge allows preimplantation genetic diagnosis (PGD) and enables prenatal DNA testing in future pregnancies. Although simultaneous testing of all FA genes by next generation sequencing will be possible in the near future, this technique will not be available immediately for all laboratories. In addition, in populations with strong founder mutations, a limited test using Sanger sequencing and MLPA will be a cost-effective alternative. We describe a strategy and optimized conditions for the screening of FANCA, FANCB, FANCC, FANCE, FANCF, and FANCG and present the results obtained in a cohort of 54 patients referred to our diagnostic service since 2008. In addition, the follow up with respect to genetic counseling and carrier screening in the families is discussed.http://dx.doi.org/10.1155/2012/603253 |
| spellingShingle | Johan J. P. Gille Karijn Floor Lianne Kerkhoven Najim Ameziane Hans Joenje Johan P. de Winter Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing Anemia |
| title | Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing |
| title_full | Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing |
| title_fullStr | Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing |
| title_full_unstemmed | Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing |
| title_short | Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing |
| title_sort | diagnosis of fanconi anemia mutation analysis by multiplex ligation dependent probe amplification and pcr based sanger sequencing |
| url | http://dx.doi.org/10.1155/2012/603253 |
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